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Figure 5


Fig. 5. P2Y2R-{alpha}v integrin interaction is required for G12 coupling. (A,B) Membrane preparations from 1321N1 cells expressing the WT or RGE mutant P2Y2R or pLXSN vector-transfected cells (negative control) were used in [35S]GTP{gamma}S binding assays in the presence or absence of 1 mM UTP. After termination of the assay, samples were immunoprecipitated with antiserum against (A) G{alpha}12 or (B) G{alpha}q/11 and radioactivity in the immunoprecipitates was calculated. Data are the means ± s.e.m. of results from three separate experiments and are shown as fold increase over untreated cells. (C) Human 1321N1 cells expressing the WT or RGE mutant P2Y2R were incubated with 1 mM UTP for 2 minutes. G12 activation was detected by immunoprecipitation (IP) of G{alpha}12 with anti-G{alpha}12 antibody and immunoblotting (IB) of G{alpha}12 with anti-phosphoserine/threonine antibody. Gq/11 activation was detected by IP with anti-phosphotyrosine antibody and IB with anti-G{alpha}q/11 antibody. Blots representative of three experiments are shown.





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